Vol.4, No.1, 27-33 (2013)
http://dx.doi.org/10.4236/as.2013.41005
Agricultural Sciences
Supplementation at casing to improve yield and
quality of white button mushroom
Yaqvob Mami1*, Gholamali Peyvast1, Mahmood Ghasemnezhad1, Farhood Ziaie2
1
Department of Horticulture Science, Faculty of Agriculture, University of Guilan, Rasht, Iran;
Corresponding Author: mami_yad_1658@yahoo.com
2
Agricultural, Medical and Industrial Research School, Nuclear Science and Technology Research Institute, Karaj, Iran
*
Received 6 October 2012; revised 13 November 2012; accepted 10 December 2012
ABSTRACT
Supplementation of substrate at casing to increase the yield and quality of mushroom [Agaricus bisporus (Lange) Sing] is an important
practice in commercial production of white button mushroom. This project was done to study
the effects of supplementing the compost at
casing with ground corn and soybean seed applied at: 0 g as control, 17, 34 and 51 g per 17 kg
compost on production and harvest quality of A.
bisporus. There were significant differences
between supplemented and non-supplemented
substrates. The 34 g soybean and 51 g corn
treatments had the highest yield. There were significant differences in quality indices of mushroom due to the type and amount of the supplement. The 51 g soybean supplement produced higher protein compared with other substrates. The highest vitamin C, total phenol, total
soluble solids and antioxidant capacity obtained
of 34 and 51 g soybeans or 34 and 51 g corn,
respectively. Increased production and quality
of A. bisporus may be achieved by addition of
suitable amounts of supplements.
Keywords: Agaricus bisporus; Compost; Corn and
Soybean
1. INTRODUCTION
The button mushroom [Agaricus bisporus (Lange)
Sing] is the most widely cultivated and consumed mushroom throughout the world and includes about 40% of
total world mushroom production [1]. The production
and the use of button mushrooms as fresh food have
newly increased in Iran. The button mushroom will grow
on composted substrates made from various materials [2].
The compost in which it grows vegetatively, and the nutritionally poor casing materials, provides suitable physical, chemical and biological conditions that stimulate
Copyright © 2013 SciRes.
initiation of fruiting body formation [3].
The casing layer, applied 14 - 16 days after spawning
is an essential part of the total substrate in the artificial
culture of A. bisporus. Although many different materials
may function as a casing layer, peat is generally regarded
as the most suitable. Because of its unique water holding
and structural properties, it is widely accepted as an ideal
for casing. Peat has a neutral pH and because of its organic content and granular structure, stays porous even
after a succession of watering, holds moisture, allows appropriate gaseous exchanges and supports microbial population able to release hormone-like substances which
are likely involved in stimulating the initiation of fruit
bodies [4,5].
Increasing the nutritional quality of mushroom compost is a prime factor in increasing yield. The previous
work showed that when compost was supplemented with
various ground seed together with refined and crude seed
oils and applied to the casing layer caused increases
mushroom yield [6]. The increase in the number and the
yield of mushrooms has also been reported when certain
chemicals were sprayed as nutrient supplementation at
casing [7]. Protein-rich supplements such as soybean,
black beans and cowpeas added at casing significantly
stimulated mushroom yield, shortened the cultivation
life-cycle, and caused colonization of the substrate compared with the non-supplemented control. Biological efficiency (BE) varied from 26.1% on unsupplemented substrate to 73.1% on compost supplemented with ground
soybean. There were no significant differences in mushroom yield observed among supplements evaluated [8].
Advantages of supplementing compost with protein, and/
or lipid-rich materials, such as corn (Zea mays L.) or
soybean (Glycinemax L.) at spawning, casing, and later
have been demonstrated [9]. Schisler, [10] reported that
supplementation of mushroom compost at spawning and
at casing with various refined and crude seed oils resulted in 0.45 to 0.68 kg–1 increases in mushroom yield.
It was determined that the ground seed or protein-oil
combinations improved sporophore yield up to 0.9 - 1.13
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28
Y. Mami et al. / Agricultural Sciences 4 (2013) 27-33
kg–1. There could be a relationship between lipid metabolism and initiation of fruiting in cultivated mushroom.
The reason may be behind the fact that sterols are involved in stimulating fruiting. Heating and stimulation of
competitor molds present in the compost limited the rate
of supplementation and benefits.
To circumvent this competition supplementation was
tried at casing [11]. Vijay et al. [12] found that supplementation at spawning or casing increased the yield of
the white button mushroom. However, it was more beneficial at casing. Gerrits, [13] found that supplementation
at casing was better than at spawning. This is probably
due to extra nutrition provided by supplements at this
stage which is directly utilized by mushroom mycelia for
increased yield. Supplementation at spawning is generally associated with a rise in temperature and incidence
of weed moulds, which may jeopardize the yield [14,15].
Improved yield and BE of Pleurotus eryngii (D.C. ex Fr.)
Qué, were achieved by supplementation of substrate with
a commercial delayed-release nutrient and the use of
casing [16].
Yield increases of 14% were achieved from cased substrates that were supplemented at time of casing with
delayed-release nutrient. The use of a casing layer enhanced the yield by 141% over non-cased substrates.
When casing and substrate supplementation were combined, the yield increased 179% over no cased/non-supplemented substrates [16]. For A. bisporus, yields for
each successive break compost decrease mainly as a consequence of nutrient depletion in the medium [17]. Supplementation at spawning or casing with delayed-release
nutrients is a practice commonly used in the commercial
cultivation of A. bisporus. The effect of some nutrient
supplementation in the casing material on crop productivity was studied by Kassim et al. [18]. The number and
yield of mushrooms increased when leucine, sodium
chloride and yeast extract were added to the casing at 10
µg/liter at three day intervals for seven weeks. Fruiting
body size was too large to check. The best yield was obtained when the trays were treated with yeast extract. The
number of days required for the appearance of primordial
fruiting bodies primordia and for the harvest was reduced
by 4 - 5 days. The aim of this study is to evaluate the effect of ground corn and soybean seed as rich sources of
protein and oil applied to casing on production and quality of white button mushroom.
2. MATERIALS AND METHODS
The experiments were conducted in the Agricultural
Faculty of Guilan University; Rasht, Iran, in 2011-2012.
The experiment was set up in a completely randomized
design with 3 replications.
Copyright © 2013 SciRes.
2.1. Compost, Casing Soil and Supplement
Preparation
Compost bags (60 × 40 × 10 cm) containing 17 kg of
compost derived from wheat straw and peat (as casing
soil) were purchased locally (Bibi, Karaj, Iran). Compost
was analyzed for chemical properties prior to use (Table
1). Corn and soybean seeds, used as supplements to the
compost, were purchased locally. Seed were ground (0.5
mm screen) and autoclaved in bags (high density polyethylene bags, 30 - 40 cm) at 110˚C (pressure 70 kPa) for
15 min. This material was mixed with peat at 0, 17, 34
and 51 g/17 kg compost prior to application as casing.
Supplements were analyzed for nitrogen amounts prior to
use (Table 2).
2.2. Adjusting the Environmental Conditions
The optimum conditions provided for spawn run in a
room were 23˚C ± 1˚C (air), RH 90% and a CO2 level of
12,000 - 15,000 µg/liter. The temperature at case run, one
stage before crop initiation (up to 1 week after casing)
was 24˚C ± 1˚C (air), RH 95% and CO2 level of 7500
µg/liter, and after an additional week it changed to 15˚C 17˚C (air), RH to 85% and CO2 level of 800 - 1000 µg/
liter by providing 6 air changes of 10 - 15 minutes/day.
2.3. Casing
After completion of mycelium running, the open top
of the spawn bags were covered with 4 cm of pasteurized
casing (peat + supplement). Watering after casing was
done as suggested for commercial growth [19,20]. The
temperature and relative humidity were 24˚C ± 1˚C and
95%, respectively in this stage.
2.4. Cropping and Harvesting
After completion of mycelium running in the casing
soil, bags were cooled to 16˚C and maintained at this
Table 1. Compost analysis.
Organic matter
71%
NH4
0.007%
C
40.5%
N
2.3%
Carbon/Nitrogen
17.6%
pH
6.8
Table 2. Supplement nitrogen content.
Supplement
N%
Corn
4.05
Soybean
7.76
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Y. Mami et al. / Agricultural Sciences 4 (2013) 27-33
29
TSS is an index of soluble solids concentration. Homogenated ground sporophore tissue was filtered through
the filter paper Whatman No. 1 by means of vacuum and
the total soluble solid of the flow throughout the process
was determined by a digital refractometer ATAGO PR32α (ATAGO, USA Inc., Kirkland, WA). The results were
expressed as Brix.
lyzed on the base of determination of free radical-scavenging effect of antioxidants on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical according to the procedure described by Elez-Martínez and Martín-Belloso [24]. Mushroom samples were centrifuged for 15 min at 4˚C (Beckman Instruments Inc., Fullerton, California) and filtered
through Whatman No. 1 filter paper. Aliquots of 0.01 mL
of the supernatant were mixed with 3.9 mL of methanolic
DPPH solution and 0.09 mL of distilled water. The homogenate was shaken vigorously and kept in the dark for
30 min. The absorption of the samples was measured
with a CECIL CE 2021 spectrophotometer (Cecil Instruments Ltd., Cambridge, UK) at 515 nm for DPPH. The
percentage of inhibition of the absorbance was calculated
and plotted as a function of the concentration of Trolox
for the standard reference data. The final DPPH value
was calculated by using a calibration curve and the percentage of inhibition was determined.
2.7. Vitamin C Content
2.11. Statistical Analysis
Vitamin C was determined by the 2,6-dichlorophenolindophenol titration method, in which the dye is reduced by
ascorbic acid, resulting in disappearance of the color [21].
The data were subjected to Analysis of Variance in
SAS (ver. 9, SAS, Inc., Cary, NC) and the means were
separated using Tukey’s test.
2.8. Protein Content
3. RESULTS
temperature and a relative humidity above 80%. Sporophores were harvested before the fruiting body showed
veil opening of the cap from the stipe. The yields of other
parameters were recorded.
2.5. Dry Matter Content
Sporophores were dried at 100˚C to determine their
dry matter content.
2.6. Total Soluble Solid (TSS)
Protein content was determined by the method of
Bradford, [22] using bovine serum albumin as the standard.
2.9. Total Phenolic Compounds
Amounts of total phenolic compounds in fresh mushrooms were determined according to the Folin-Ciocalteu
procedure [23] with some modifications. For this purpose, 0.5 g mushroom slices was ground in the presence
of liquid nitrogen by mortar and pestle, then 10 mL pure
methanol were added to it for extracting phenolic compounds. The extract was centrifuged with 3000 rpm for
15 min at 4˚C with an AVANTI™ J-25 centrifuge (Beckman Instruments Inc., Fullerton, CA) and then filtered
through a Whatman No. 1 filter paper. Then 300 µl of
methanolic extract was brought to a volume of 500 µl
with distilled water into test tubes, followed by 2.5 mL of
10% Folin-Ciocalteu reagent and allowed to stand for 6
min. Thereafter, 2 mL of 7.5% sodium carbonate solution
were added. Each sample was allowed to stand for 90
min at room temperature in darkness and the absorbance
was measured at 760 nm using an UV/Vis spectrophotometer model PG Instrument + 80 (Leicester, UK). Results are expressed as mg gallic acid/100 g fresh weight.
Each assay was carried out in triplicate.
2.10. Antioxidant Capacity
The antioxidant capacity of fresh mushroom was anaCopyright © 2013 SciRes.
Our results indicated that different levels of supplements affected the yield and the harvest quality. Yield,
vitamin C, TSS, protein, antioxidant capacity and total
phenol were the lowest, when the compost was not
treated with the additives. Soybean at 34 g produced better yield, fruit number (small, average and total) and dry
matter compared with other treatments. Soybean (34 g)
and corn (51 g) were not different (Figures 1-5). The
highest amounts of TSS and total phenol were produced
by adding the corn (51 g). TSS and total phenol content
did not differ significantly with CR 34 g and CR 51 g
(Figures 6 and 10). Most Vitamin C content; was obtained from the soybean (34 and 51 g). The highest protein and antioxidant capacities (AC) in mushrooms were
due to the use of soybean (51 g) and corn (34 g), respectively. AC did not differ significantly with CR 34 g and
CR 51 g (Figures 7-9).
4. DISCUSSION
Application of a casing layer for A. bisporus production is essential. Casing layer depth, chemical and microbial composition, physical properties and moisture
content of the casing layer play important roles in yield
and quality of mushrooms [25-30]. Limited information
is available regarding the usage of supplements at casing
versus at spawning for improvement of production and
mushroom quality. Supplementing substrate at casing is a
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30
Y. Mami et al. / Agricultural Sciences 4 (2013) 27-33
a
Yield (kg/m2)
20
cd
ab
cb
d
cd
cd
cd
Dry matter (%)
25
15
10
5
20
18
16
14
12
10
8
6
4
2
0
a
0
C1
C2
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
cab
cb
c
7
6
ab
cab
cab
d
d
C1
C2
5
b
dcb
dc
ab
cb
a
4
3
2
1
0
600
a
cab
500
c
ab
cb
ab
cb
cab
300
200
100
Supplement (g)
Figure 6. Effect of supplements in case soil on total solid
soluble in A. bisporus. C1 (Control for soybean), C2
(Control for corn), SB (Soybean) and CR (Corn) (n = 3).
Vit C.
( m g / 100g f. wt)
Figure 2. Effect of supplements in case soil on fruit number (small) in A. bisporus. C1 (Control for soybean), C2
(Control for corn), SB (Soybean) and CR (Corn) (n = 3).
400
4
3.5
3
2.5
2
1.5
1
0.5
0
0
a
c
C1
C1
C2
C1
cb
C2
c
c
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
Figure 4. Effect of supplements in case soil on total fruit
number in A. bisporus. C1 (Control for soybean), C2
(Control for corn), SB (Soybean) and CR (Corn) (n = 3).
Copyright © 2013 SciRes.
b
b
Supplement (g)
Figure 7. Effect of supplements in case soil on vit C. in A.
bisporus. C1 (Control for soybean), C2 (Control for corn),
SB (Soybean) and CR (Corn) (n = 3).
ab
ab
bc
C2 SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
a
c
c
a
bc
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Figure 3. Effect of supplements in case soil on fruit number (averege) in A. bisporus. C1 (Control for soybean), C2
(Control for corn), SB (Soybean) and CR (Corn) (n = 3).
c
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
C2 SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
fruit number
(1.5 ≤ cap ≤ 4)/m2
C2
c
Figure 5. Effect of supplements in case soil on dry matter
in A. bisporus. C1 (Control for soybean), C2 (Control for
corn), SB (Soybean) and CR (Corn) (n = 3).
TSS (°Brix)
a
cab
C1
Total fruit number/m2
C1
b
Supplement (g)
Protein
(g / 100g f. wt)
Fruit number
(cap<1.5cm)/m2
Figure 1. Effect of supplements in case soil on yield in A.
bisporus. C1 (Control for soybean), C2 (Control for corn),
SB (Soybean) and CR (Corn) (n = 3).
1000
900
800
700
600
500
400
300
200
100
0
d
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
450
400
350
300
250
200
150
100
50
0
d
a
a
b
3.5
3
2.5
2
1.5
1
0.5
0
a
d
C1
d
c
b
d
d
c
C2 SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
Figure 8. Effect of supplements in case soil on protein in
A. bisporus. C1 (Control for soybean), C2 (Control for
corn), SB (Soybean) and CR (Corn) (n = 3).
OPEN ACCESS
AC
(mmol/kg f wt)
Y. Mami et al. / Agricultural Sciences 4 (2013) 27-33
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
b
d
C1
d
a
c
ab
c
d
C2 SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (gr)
Figure 9. Effect of supplements in case soil on antioxidant capacity in A. bisporus. C1 (Control for soybean),
C2 (Control for corn), SB (Soybean) and CR (Corn) (n =
3).
Total phenol
(m g / 100g f. wt )
250
b
200
150
c
c
C1
C2
a
b
a
b
c
100
50
0
SB 17 SB 34 SB 51 CR 17 CR 34 CR 51
Supplement (g)
Figure 10. Effect of supplements in case soil on total
phenol in A. bisporus. C1 (Control for soybean), C2
(Control for corn), SB (Soybean) and CR (Corn) (n = 3).
relatively easy and low-cost cultural practice that may
successfully be used to enhance the yield, BE and maximize the utilization of substrate. Since microbial populations in casing material are important in promoting fructification in A. bisporus, supplementing at casing can
also affect microorganism populations. A direct correlation between the mushroom antioxidant capacity and total phenolic content had been reported in the light of the
antioxidant action raised by other substances such as tocopherols and β-carotene [31-33]. However, increased
antioxidant capacity after the supplementation with soybean could be also related to high content of polyphenolic isoflavonoids such as genistein, genistin, daidzein,
daidzin, formononetin, and afrormosin in soybean [34].
Vijay et al. [12] indicated that supplementation at spawning, or at casing, significantly increased the yield. However, it was more beneficial at casing. Gerrits [13] found
that supplementation at casing was better than at spawning. This is probably due to the fact that extra nutrition
was provided by the supplements at this stage which is
directly utilized by mycelia. Contrary to the casing process supplementation at spawning is generally associated
with a rise in temperature and incidence of weed moulds,
which may jeopardize the yield. Similar results have
been reported [13,14,35]. In lipid metabolism, studies
Copyright © 2013 SciRes.
31
concerned with A. bisporuss howed that the mushroom
yield was increased by the use of seed oils [7,36]. Carbon supplied to the compost in oil supplementation at
casing was utilized to synthesize mannitol and fatty acids
[37]. Mannitol is the single largest component of the dry
matter in the mushroom comprising as much as 19% of
dry weight. Mannitol provides turgor pressure for support of the fruit body and for the considerable extension
growth which occurs [38]. Due to our results, supplementation with ground corn and soybean seeds increased
the yield and quality of A. bisporus more than control
treatment. Consequently, we can recommend the supplementation with corn and soybean seeds at casing as a
good practice to increase the production and mushroom
quality.
5. CONCLUSION
We found that yield and quality of mushroom can be
improved by using the supplements at casing soil. This
experiment also, showed that treatments of soy bean 34
and corn 51 induced better yields compared with other
treatments. In addition, amounts of quality indices in
mushrooms were different among supplemented composts with soybean and corn.
6. ACKNOWLEDGEMENTS
Authors acknowledge the support of University of Guilan, Rasht, Iran.
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